Journal of Arak University of Medical Sciences
مجله دانشگاه علوم پزشكي اراك
J Arak Uni Med Sci
Medical Sciences
http://jams.arakmu.ac.ir
46
journal46
1735-5338
2008-644X
fa
jalali
1398
3
1
gregorian
2019
6
1
22
2
online
1
fulltext
fa
بررسی اثر داروی سیسپلاتین و عصاره اوکالیپتوس بر روی بیان ژن KRAS در رده سلولی A549 سرطان ریه
The Effect of Cisplatin and Eucalyptus Extract on KRAS Gene Expression in A549 Cell Line of Lung Cancer
علوم پایه
Basic Sciences
پژوهشي اصیل
Original Atricle
<div style="text-align: justify;"><strong><span style="color:#12336E;"><span style="font-family:b nazanin;">زمینه و هدف</span></span></strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">: سرطان ریه یکی از همه</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">گیرترین سرطان</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">ها در سراسر جهان است. اخیراً چند انکوژن بالقوه شناخته شده</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">اند که شامل </span></span><span dir="LTR"><span style="font-family:times new roman,serif;">EGFR</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span><span dir="LTR"><span style="font-family:times new roman,serif;">BRAF</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">، </span></span><span dir="LTR"><span style="font-family:times new roman,serif;">KRAS</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> و ژن</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> ALK </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">هستند. با توجه به شیوع بالای سرطان ریه، میزان مرگ ناشی از آن، عوارض شیمی</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">درمانی و تلاش برای یافتن داروهای مؤثر و با عوارض کمتر، تحقیق حاضر با این هدف انجام شد که تأثیر یک عصاره گیاهی را بررسی کرده تا نتایج آن در اختیار مراکز تولیدی قرار گیرد.</span></span><br>
<strong><span style="color:#12336E;"><span style="font-family:b nazanin;">مواد و روش</span></span></strong><strong><span dir="LTR"><span style="color:#12336E;"><span style="font-family:times new roman,serif;"></span></span></span></strong><strong><span style="color:#12336E;"><span style="font-family:b nazanin;">ها:</span></span></strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> در این تحقیق، تأثیر عصاره اوکالیپتوس و داروی سیس</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">پلاتین بر روی میزان بیان ژن</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> KRAS </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">در رده سلولی</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> A549 </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">سرطان ریه بررسی شد. به منظور تعیین زنده</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">مانی سلول</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">ها از روش</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> MTT </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">استفاده و</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> IC50 </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">تعیین شد. پس از تعیین </span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> IC50</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">سلول</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">ها به مدت 48 ساعت در معرض تأثیر غلظتی کمتر از</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> IC50 </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">عصاره و دارو قرار گرفتند. سپس میزان بیان ژن</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> KRAS </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">نسبت به ژن کنترل</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> β-ACTIN </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">در گروه</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">های کنترل و تیمار با عصاره و دارو تعیین شد. بدین منظور، پرایمر اختصاصی برای ژن</span></span><span dir="LTR"><span style="font-family:times new roman,serif;">β-ACTIN </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> و</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> KRAS </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">طراحی و</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> Real-Time PCR</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> انجام شد.</span></span><br>
<strong><span style="color:#12336E;"><span style="font-family:b nazanin;">ملاحظات اخلاقی: </span></span></strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">این مطالعه با کد </span></span><span dir="LTR"><span style="font-family:times new roman,serif;">IR.IAU.East Tehran.REC.1396.3</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> در کمیته اخلاق پژوهشی دانشگاه آزاد اسلامی واحد تهران شرق به تصویب رسیده است. </span></span><br>
<strong><span style="color:#12336E;"><span style="font-family:b nazanin;">یافته</span></span></strong><strong><span dir="LTR"><span style="color:#12336E;"><span style="font-family:times new roman,serif;"></span></span></span></strong><strong><span style="color:#12336E;"><span style="font-family:b nazanin;">ها:</span></span></strong><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> نتایج نشان داد میزان</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"> IC50 </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">عصاره و دارو به ترتیب</span></span> <span style="font-family:b nazanin;"><span style="font-size:12.0pt;">7/85 و 77/1 میلی</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">گرم بر میلی</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">لیتر</span></span> <span style="font-family:b nazanin;"><span style="font-size:12.0pt;">بوده است. هم</span></span><span dir="LTR"><span style="font-family:times new roman,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">چنین، میزان بیان ژن</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">ها در سلول</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">های تیمار شده و کنترل با عصاره و دارو مقایسه گردید. بیان نسبی ژن</span></span><span dir="LTR"><span style="font-family:times new roman,serif;">KRAS </span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> به ژن مرجع در رده سلولی سرطانی تیمارشده با عصاره و دارو به ترتیب به میزان 89/2 و 25/9 برابر طی 48 ساعت بهطور معنی</span></span><span dir="LTR"><span style="font-family:cambria,serif;"></span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">دار (0=</span></span><span dir="LTR"><span style="font-family:times new roman,serif;">p</span></span><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">) کاهش یافت.</span></span><br>
<strong><span dir="RTL"><span style="color:#12336E;"><span style="font-family:b nazanin;"><span style="font-size:11.0pt;">نتیجه</span></span></span></span><span style="color:#12336E;"><span style="font-family:times new roman,serif;"><span style="font-size:11.0pt;"></span></span></span><span dir="RTL"><span style="color:#12336E;"><span style="font-family:b nazanin;"><span style="font-size:11.0pt;">گیری:</span></span></span></span></strong><span dir="RTL"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> با توجه به کاهش بیان نسبی ژن در رده </span></span></span><span style="font-family:times new roman,serif;"><span style="font-size:11.0pt;">A549</span></span><span dir="RTL"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;"> تیمار شده می</span></span></span><span style="font-family:cambria,serif;"><span style="font-size:11.0pt;"></span></span><span dir="RTL"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">توان به مطالعات آینده در خصوص درمان هدف</span></span></span><span style="font-family:times new roman,serif;"><span style="font-size:11.0pt;"></span></span><span dir="RTL"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">مند سرطان ریه امیدوار بود و از این</span></span></span><span style="font-family:times new roman,serif;"><span style="font-size:11.0pt;"></span></span><span dir="RTL"><span style="font-family:b nazanin;"><span style="font-size:12.0pt;">رو پتانسیل استفاده از ترکیبات گیاهی آشکارتر شود</span></span></span><span dir="RTL"><span style="font-family:b nazanin;"><span style="font-size:11.0pt;">. </span></span></span></div>
<div style="text-align: justify;"><span style="font-family:times new roman;"><strong>Background and Aim:</strong> Lung cancer is one of the most contagious cancers in all of the world. Recently, several potential oncogenes and carcinogens have been identified, including EGFR, BRAF, KRAS and ALK genes. With due attention to the high prevalence of lung cancer, its death rate, the complications of chemotherapy and the efforts to find effective and less effective drugs, this study was done to investigate the effect of a plant extract so that results are available to manufacturing centers.<br>
<strong>Materials and Methods:</strong> In this study, the effect of Eucalyptus extract and cisplatin on the expression of KRAS gene in A549 lung cancer cell line was investigated. To determine the cell survival, MTT was used and IC50 was determined. After determining IC50, the cells were exposed to less than IC50 concentrations of the extract and drug for 48 hours. Then, the amount of β-ACTIN and KRAS genes expressions in control and extract treated and drug treated groups were determined. For this purpose, a specific primers were designed for β- ACTIN and KRAS, and Real-Time PCR was be done.<br>
<strong>Ethical Considerations:</strong> This study with research ethics code IR.IAU.East Tehran.REC.1396.3 has been approved by research ethics committee at Islamic Azad University, Tehran- East Branch, Iran.<br>
<strong>Findings:</strong> The results showed that the amount of IC50 of the extract and drug was 8.75 and 1.77 mg/ml, respectively. In addition, the expression of genes in control and treated cells with extract and drug was compared. The expression of the KRAS gene relative to the reference gene in the cancer cell line treated with extract and drug, for 48 hours, was significantly decreased 2.89 and 9.25, respectively (p = 0).<br>
<strong>Conclusion: </strong>Regarding the reduction of the relative gene expression in the A549 treated group, future studies on targeted lung cancer treatment can be promising and the potential for the use of plant compounds is more evident.</span></div>
ژن KRAS, سرطان ریه, سیسپلاتین, عصاره اوکالیپتوس
Cisplatin, Eucalyptus extract , KRAS gene, Lung cancer
84
95
http://jams.arakmu.ac.ir/browse.php?a_code=A-10-5286-3&slc_lang=fa&sid=1
Mahsa
Kavousi
مهسا
کاوسی
mkavoosi@iauet.ac.ir
4600319475328460066915
4600319475328460066915
Yes
Department of Biology, East Tehran Branch, Islamic Azad University,Tehran, Iran.
گروه زیست شناسی، واحد تهران شرق، دانشگاه آزاد اسلامی، تهران، ایران.
Ehsan
Rahimi
احسان
رحیمی
rahimi_en@yahoo.com
4600319475328460066916
4600319475328460066916
No
Department of Biology, East Tehran Branch, Islamic Azad University,Tehran, Iran.
گروه زیست شناسی، واحد تهران شرق، دانشگاه آزاد اسلامی، تهران، ایران.
Jalil
Fallah Mehrabadi
جلیل
فلاح مهرآبادی
jalil.falla@gmail.com
4600319475328460066917
4600319475328460066917
No
Lister Laboratory of Microbiology, Tehran, Iran.
آزمایشگاه میکروب شناسی لیستر، تهران، ایران.